Skincare Methods

ABSTRACT

Disclosed are compositions and methods for skincare, e.g., reducing skin wrinkles and for treating skin disorders.

RELATED APPLICATIONS

This application is a continuation-in-part of U.S. patent application.No. 11/848,272, filed on Aug. 31, 2007, which is a continuation-in-partof: (A) U.S. patent application Ser. No. 11/499,936, filed on Aug. 7,2006, which is a continuation-in-part of U.S. patent application Ser.No. 10/798,119, filed on Mar. 11, 2004, which is a continuation-in-partof U.S. patent application Ser. No. 10/205,738, filed on Jul. 25, 2002(now U.S. Pat. No. 6,809,118); (B) U.S. patent application Ser. No.10/843,025, filed on May 10, 2004, which is a continuation-in-part ofU.S. patent application Ser. No. 10/205,738, filed on Jul. 25, 2002 (nowU.S. Pat. No. 6,809,118); and (C) a continuation-in-part of U.S. patentapplication Ser. No. 11/079,370, filed on Mar. 14, 2005, which is acontinuation-in-part of U.S. patent application Ser. No. 10/132,999,filed on Apr. 26, 2002. The prior applications are incorporated hereinby reference in their entirety

BACKGROUND

Skin, the largest organ of the human body, has important functions inprotecting the body from infection, toxins, microbes, and radiation.Although the type of skin varies over the body, skin generally has twomain layers of tissue. The epidermis or cuticle, the outermost layer, iscomposed of three superficial and two deep cellular layers. The derma,corium, or cutis vera, the true skin, is composed of a papillary layerabove and a reticular layer below. It contains blood vessels thatnourish the skin cells and the structural elements e.g., collagen andelastin, which keep the skin firm and springy.

The epithelial cells of the skin are constantly dying and being replacedby new cells, to the extent that human skin renews its surface layersevery three to four weeks. Unlike the epithelial cells, collagen cellsin the subcutaneous layer of will not be replaced for around 30 years.When these old collagen cells breakdown due to aging, sun damage,smoking, poor hydration or steroid overuse, the fragmented collagencauses folds, ridges or creases in the skin, i.e., wrinkles. Fragmentedcollagen also leads to skin tearing and bruising easier than itotherwise would. Wrinkles are a reflection of slower skin regeneration.The color and appearance of the skin deteriorates slowly due to aging,exposure to sunlight and radiation, and/or degradation of thedermal-epidermal junction and of the cell-cell cohesion in theepidermis. Because the changes of skin appearance cause significantlypsychological, social and occupational problems, there is an increasingdemand in mitigating or delaying the dermatological signs ofchronologically or photo-aged skin, such as fine lines, wrinkles, andsagging skin.

SUMMARY

This invention relate to a method for ameliorating skin wrinkles andrejuvenating skin.

Accordingly, one aspect of this invention features a method for reducingskin wrinkles. The method includes administering to a subject in needthereof a composition containing an effective amount of a histonehyperacetylating agent or a pharmaceutically acceptable salt thereof,and a cosmetically or pharmaceutically acceptable carrier. The inventionalso features a method for treating a skin disorder, such as acne andurticaria. The method can further include identifying a subject havingthe disorder or skin wrinkles before the administering, or, examiningthe subject for the effect of the composition on the disorder or skinwrinkles after the administering the composition.

The histone hyperacetylating agent can be a histone deacetylase (HDAC)inhibitor. Examples of the HDAC inhibitor include, but not limited to,trichostatin A, trichostatin C, oxamflatin, trapoxin A, FR901228,apicidin, HC-Toxin, WF27082, chlamydocin, salicylihydroxamic acid,suberoylanilide hydroxamic acid, azelaic bishydroxamic acid,azelaic-1-hydroxamate-9-an-ilide, M-carboxycinnamic acid bishydroxamide,6-(3-chlorophenylureido)carpoic hydroxamic acid, MW2796, MW2996, sodiumbutyrate, arginine butyrate, isovalerate, valerate, 4-phenylbutyrate,sodium phenylbutyrate, propionate, butrymide, isobutyramide,phenylacetate, 3-bromopropionate, valproic acid, valproate, tributyrin,MS-27-275 or a 3′-amino derivative thereof, depudecin and scriptaid. Thehistone hyperacetylating agent can be present in an amount of from0.00001% to 100%, e.g., 0001% to 10%, by weight of the composition.

In one embodiment, the above-mentioned composition is a topicalcomposition and the administering step is conducted by applying thecomposition to a surface of skin in need thereof of the subject. Thecomposition can be a cream, an ointment, a gel, a paste, a powder, anaqueous solution, a spray, a suspension, a dispersion, a salve, alotion, a patch, a suppository, a liposome formation, a mouth wash, anenema, an injection solution, an eye drop, an ear drop, a drip infusion,a microcapsule, or a nanocapsule. The composition can further contain apenetration enhancing agent, or a pH adjusting agent to provide aformulation pH in the range of approximately 2.0 to 13.0. It can be usedas an adjuvant treatment following laser surgery or laser dermabrasiontreatment to enhance aged and injured skin rejuvenation.

The above described method can further include administering to thesubject a second agent. This second agent can include a cytokine, acytokine antagonist, an interleukin, an interleukin antagonist, a growthfactor, an angiogenic agent, an anti-histamine, an anti-fibrogenicagent, a vasoactive agent, an antibody, a conjugated antibody, anadenosine receptor agonist, a peroxisome proliferating activatorreceptor (PPAR) agonist, an anticholinergics, a non-steroidanti-inflammation drug (NSAID), a steroid, an anti-oxidant agent, avitamin, a leukotriene modifier, a mast cell inhibitor, an anti-IgEantibody, lidocaine, epinephrine, a selective serotonin reuptakeinhibitor (SSRI), a 5-hydroxytryptamine (5-HT) receptor antagonist, anantibiotics, a calcineurin inhibitor, an amino acid, a matrixmetalloproteinase (MMP) inhibitor, a DNA methylation inhibitor,collagenase, clostridium histolyticum, or combinations thereof. Theabove-described composition and the second agent can be systemically ortopically administered simultaneously or sequentially.

The details of one or more embodiments of the invention are set forth inthe accompanying drawings and the description below. Other features,objects, and advantages of the invention will be apparent from thedescription and drawings, and from the claims.

DESCRIPTION OF DRAWINGS

FIG. 1 is a diagram showing skin recovery and accelerated effect of a 5%sodium 4-phenylbutyrate (an HDAC inhibitor) gel (YLC-Gel) on skinregeneration after abrasive damage in a mouse model.

FIGS. 2A-2L are H&E histology photographs showing that topical sodium4-phenylbutyrate (PB), an HDAC inhibitor, has effects of amelioratingwrinkles, producing smooth skin and rejuvenating skin after radiationdamage. FIGS. 2A, 2D, 2G, and 2J are H&E histology at 40× field; 2B, 2E,2H, and 2K are H&E histology at 100× field; 2C, 2F, 2I, and 2L are H&Ehistology at 200× field. FIGS. 2A-2C are of normal skin. FIGS. 2D-2F areof acute reaction on Day 7 after irradiation, showing subepithelialswelling (black arrow). FIGS. 2G-2I are of the vehicle group on Day 180,showing wrinkles and thick dermis with more fragmented collagen deposit,thinner epithelium, subepithelium swelling, and increased vessels. Theblack arrows indicate that the subcutaneous fat layer in the vehiclegroup was replaced by more fragmented collagen tissues. FIGS. 2J-2L areof phenylbutyrate (PB) treated group on Day 180, showing smooth skin andthinner dermis with less fragmented collagen, thicker epidermis with10-30 cell layers (black arrow), and less subepithelial swelling. InFIG. 2A, a indicates epidermis, b indicates dermis, and c indicatessubcutaneous tissue.

FIGS. 3A-3D are photographs of immunofluorescence with the anti-TGF-beta1, 2 antibodies showing that the expression of TGF-beta, a fibrogenicgrowth factor, was suppressed by phenylbutyrate (PB) as shown in Example3 below. FIG. 3A is of normal skin stained with TGF-beta. FIG. 3B is apicture of acute dermatitis on Day 7 after irradiation without any drugtreatment, showing that TGF-beta was up-regulated. FIG. 3C is of thevehicle group on Day 180 after irradiation, showing that the expressionof TGF-beta was increased with time, persistent and highly expressed infibrogenic skin wrinkles both in keratinocytes of the epidermis and inmyofibroblasts of the dermis with more fragmented collagen deposit. FIG.3D is of the phenylbutyrate (PB) treated-group on Day 180 afterirradiation, showing that the topical phenylbutyrate (PB) suppressed theTGF-beta expression effectively, which correlates well with lessfragmented collagen fiber accumulation in the dermis and more celllayers in the epithelium. In FIGS. 3A and D, a indicates epidermis, bindicates dermis, and c indicates subcutaneous tissue.

FIGS. 4A-4D are photographs of immunohistochemistry with theanti-TNF-alpha antibody showing that the expression of TNF-alpha, aproinflammatory cytokine, was suppressed by phenylbutyrate (PB). FIG. 4Ais of normal skin for TNF-alpha staining FIG. 4B is of thephenylbutyrate (PB) treated-group on Day 270 after irradiation, showingthat phenylbutyrate (PB) suppressed the TNF-alpha expressioneffectively, which correlates well with less inflammatory cellinfiltration and no chronic ulceration. FIG. 4C is of an irradiated skintreated with Vaseline, showing that TNF-alpha was up-regulated in thesubcutaneous tissue with ulcerations on Day 270 (the arrow indicates thenecrotic wound). FIG. 4D is an irradiated skin treated with vehicle,showing that TNF-alpha was up-regulated in the subcutaneous tissue withheavy inflammatory cell infiltrates on Day 270. In FIG. 4B, b indicatesdermis, and c indicates subcutaneous tissue.

FIGS. 5A-5B are photographs showing that the topical 10% phenylbutyratecream rejuvenates skin from bacterial injection, inflammation and immunereaction. FIG. 5A is a photograph of a paw treated with phenylbutyrate,which promotes the wound healing and produces smooth skin in thebacterial injection site in the plantar region; FIG. 5B is a photographof a paw treated with vehicle (white arrow indicates the ulcer resultingfrom the injection of killed Mycobacterium tuberculosis with 0.3 mg in0.1 ml of light mineral oil and complete Freund's Adjuvant).

FIGS. 6A and 6B are a diagram and a set of photographs showing that thetopical 2.5% phenylbutyrate gel suppresses the tyrosine kinase inhibitor(TKI)-induced skin reaction in a mouse model. TKI induces acneform-likeskin lesions in humans, and augments skin swelling in mice. FIG. 6A is atime-course diagram demonstrating phenylbutyrate can ameliorate the TKI(PD168393)-induced skin toxicity in mice. FIG. 6B shows the comparisonof H&E histology of representative examples of the TKI (PD168393)-augmented skin reaction to DNFB irritation at 48 hours in theears of mice treated with or without the placebo gel or 2.5%phenylbutyrate gel.

DETAILED DESCRIPTION

This invention is based, at least in part, on unexpected findings that acomposition containing a histone hyperacetylating agent regulates andimproves skin conditions. The present invention therefore features amethod of using a histone hyperacetylating agent, e.g., a histonedeacetylase inhibitor, for ameliorating wrinkles and rejuvenating skin.The method disclosed in this invention is particularly useful forimproving aesthetic appearance of skin and for producing smooth skin.

Histone Deacetylase

Histone deacetylases (HDACs) refer to enzymes that catalyze the removalof acetyl groups from lysine residues in the amino terminal tails of thenucleosomal core histones. As such, HDACs together with histone acetyltransferases (HATs) regulate the acetylation status of histones. Histoneacetylation affects gene expression and inhibitors of HDACs, such as thehydroxamic acid-based hybrid polar compound suberoylanilide hydroxamicacid (SAHA) induce growth arrest, differentiation and/or apoptosis oftransformed cells in vitro and inhibit tumor growth in vivo. HDACs canbe divided into three classes based on structural homology. Class IHDACs (HDACs 1, 2, 3 and 8) bear similarity to the yeast RPD3 protein,are located in the nucleus and are found in complexes associated withtranscriptional co-repressors. Class II HDACs (HDACs 4, 5, 6, 7 and 9)are similar to the yeast HDA1 protein, and have both nuclear andcytoplasmic subcellular localization. Both Class I and II HDACs areinhibited by hydroxamic acid-based HDAC inhibitors, such as SAHA. ClassIII HDACs form a structurally distant class of nicotinamide (NAD)dependent enzymes that are related to the yeast SIR2 proteins and arenot inhibited by hydroxamic acid-based HDAC inhibitors.

Histone Deacetylase (HDAC) Inhibitors

HDAC inhibitors, as used herein, refer to compounds that are capable ofinhibiting the deacetylation of histones in vivo, in vitro or both. Assuch, HDAC inhibitors inhibit the activity of at least one histonedeacetylase. As a result of inhibiting the deacetylation of at least onehistone, an increase in acetylated histone occurs and accumulation ofacetylated histone is a suitable biological marker for assessing theactivity of HDAC inhibitors. Therefore, procedures which can assay forthe accumulation of acetylated histones can be used to determine theHDAC inhibitory activity of compounds of interest. It is understood thatcompounds which can inhibit histone deacetylase activity can also bindto other substrates and as such can inhibit other biologically activemolecules such as enzymes or non-histone proteins such astranscriptional factors, heat-shock proteins, chaperones and structuralproteins.

HDAC inhibitors as a class of compounds with abilities in multiple generegulation can modulate the expression of a specific set of genes byincreasing histone acetylation, thereby regulating chromatin structureand accessibility of target genes for transcription and thus treatingdiseases (Marks, P A., et al., J. Natl. Cancer Inst., 92: 1210-6, 2000).HDAC inhibitors act selectively on gene expression, altering theexpression of only about 2% of the genes expressed in cultured tumorcells. By modulating specific genes related to cell cycle control, DNArepair, tumor suppression, apoptosis and oncogenesis, HDAC inhibitorshave shown to be potent inducers of growth arrest, differentiation,and/or apoptotic cell death of transformed cells in vitro and in vivo.Although the effects of HDAC inhibitors induce bulk histone acetylation,they result in apoptotic cell death, terminal differentiation, andgrowth arrest only in tumor cells but no toxicity in normal cells(Richon, V M., et al., Proc. Natl. Acad. Sci. USA., 97: 10014-10019,2000; Van Lint, C., et al., Gene Expr., 5: 245-243, 1996). Theepigenetic modification of chromatin structure for gene regulationsuggests that HDAC inhibitors could be therapeutic candidates not onlyfor cancers but also for genetic disorders such as cystic fibrosis,sickle cell anemia, beta-thalassemia, X-linked adrenoleukodystrophy,spinal muscular atrophy, and neurodegenerative disorders, and aging(Kemp S, et al. Nat Med 4: 1261-8, 1998; et al. Proc Natl Acad Sci USA102: 11023-8, 2005; Kang H L, et al. Proc Natl Acad Sci USA 99: 838-43,2002). On the other hand, HDAC inhibitors can also induce non-histoneprotein hyperacetylation. The hyperacetylation of nonhistone proteinssuch as ribosomal S3 or the Rel-A subunit of NF-kappaB will inhibit theNF-kappaB activity and suppress the pro-inflammatory cytokine production(Chen, L., et al., Science, 293: 1653-1657, 2001; Blanchard F, et al.Drug Discov Today 10: 197-204, 2005). Thus, in addition to theanti-cancer and gene modulation effects, HDAC inhibitors have alsodemonstrated anti-inflammatory effects on many inflammation diseasessuch as ulcerative colitis and autoimmune diseases (Segain, J P., etal., Gut, 47: 397403, 2000; Mishra, N., et al., Proc. Natl. Acad. Sci.USA., 98: 2628-2633, 2001; Leoni, F., et al., Proc. Natl. Acad. Sci.USA, 99: 2995-3000, 2002; Chung, Y L., et al., Mol. Ther. 8: 707-717,2003).

On the basis of the abilities in coordinately, selectively,differentially and epigenetically modulating the expression of cellgrowth control genes, proinflammatory cytokines (IL-1, TNF-alpha), andfibrogenic growth factors (TGF-beta), a pharmaceutical compositioncomprising the HDAC inhibitor may provide an effective treatment notonly to accelerate wound healing but also to prevent scar formation,thus resulting in less fragmented collagen accumulation in the dermis toameliorate wrinkle formation and produce smooth skin.

This invention discloses a method for skin rejuvenation by using HDACinhibitors to protect skin cells against various physiological and/orpathological stresses, and to produce youthful-appearing skin. Activecompounds used to carry out the present invention are, in general,hyperacetylating agents, such as HDAC inhibitors. Numerous suchcompounds are known. See, e.g., P. Dulski, Histone Deacetylase as Targetfor Antiprotozoal Agents, PCT Application WO 97/11366 (Mar. 27, 1997).Examples of such compounds include, but are not limited to:

A. Trichostatin A and its analogues such as: trichostatin A (TSA); andtrichostatin C (Koghe et al. 1998. Biochem. Pharmacol. 56:1359-1364)(Trichostatin B has been isolated but not shown to be an HDACinhibitor).

B. Peptides, such as: oxamflatin [(2E)-5-[3-[(phenylsulfonyl)aminophenyl]-pent-2-en-4-ynohydroxamic acid (Kim et al. Oncogene,18:2461-2470 (1999)); trapoxin A (TPX)-Cyclic Tetrapeptide(cyclo-(L-phenylalanyl-L-phenylalanyl-D-pipecolinyl-L-2-amino-8-oxo-9,10-epoxy-decanoyl))(Kijima et al., J. Biol. Chem. 268, 22429-22435 (1993)); FR901228,depsipeptide (Nakajima et al., Ex. Cell Res. 241, 126-133 (1998));FR225497, cyclic tetrapeptide (H. Mori et al., PCT Application WO00/08048 (Feb. 17, 2000)); apicidin, cyclic tetrapeptide[cyclo(N-O-methyl-L-tryptophanyl-L-isoleucinyl-D-pipecolinyl-L-2-amino-8-oxodecanoyl)-](Darkin-Rattray et al., Proc. Natl. Acad. Sci. USA 93, 13143-13147(1996)); apicidin Ia, apicidin Ib, apicidin Ic, apicidin IIa, andapicidin IIb (P. Dulski et al., PCT Application WO 97/11366); HC-Toxin,cyclic tetrapeptide (Bosch et al., Plant Cell 7, 1941-1950 (1995));WF27082, cyclic tetrapeptide (PCT Application WO 98/48825); andchlamydocin (Bosch et al., supra).

C. Hydroxamic acid-based hybrid polar compounds (HPCs), such as:salicylihydroxamic acid (SBHA) (Andrews et al., International J.Parasitology 30, 761-768 (2000)); suberoylanilide hydroxamic acid (SAHA)(Richon et al., Proc. Natl. Acad. Sci. USA 95, 3003-3007 (1998));azelaic bishydroxamic acid (ABHA) (Andrews et al., supra);azelaic-1-hydroxamate-9-anilide (AAHA) (Qiu et al., Mol. Biol. Cell 11,2069-2083 (2000)); M-carboxycinnamic acid bishydroxamide (CBHA) (Riconet al., supra); 6-(3-chlorophenylureido) carpoic hydroxamic acid(3-C1-UCHA) (Richon et al., supra); MW2796 (Andrews et al., supra); andMW2996 (Andrews et al., supra). Note that analogs not effective as HDACInhibitors are: hexamethylene bisacetamide (HBMA) (Richon et al. 1998,PNAS, 95:3003-3007); and diethylbis(pentamethylene-N,N-dimethylcarboxamide) malonate (EMBA) (Richon etal. 1998, PNAS, 95:3003-3007).

D. Short chain fatty acid (SCFA) compounds, such as: sodium butyrate(Cousens et al., J. Biol. Chem. 254, 1716-1723 (1979)); isovalerate(McBain et al., Biochem. Pharm. 53:1357-1368 (1997)); valproic acid;valerate (McBain et al., supra); 4-phenylbutyrate (4-PBA) (Lea andTulsyan, Anticancer Research, 15, 879-873 (1995)); phenylbutyrate (PB)(Wang et al., Cancer Research, 59, 2766-2799 (1999)); propionate (McBainet al., supra); butrymide (Lea and Tulsyan, supra); isobutyramide (Leaand Tulsyan, supra); phenylacetate (Lea and Tulsyan, supra);3-bromopropionate (Lea and Tulsyan, supra); and tributyrin (Guan et al.,Cancer Research, 60, 749-755 (2000)).

E. Benzamide derivatives, such as: MS-27-275[N-(2-aminophenyl)-4-[N-(pyridin-3-yl-methoxycarbonyl)aminomethyl]benzamide](Saito et al., Proc. Natl. Acad. Sci. USA 96, 4592-4597 (1999)); and3′-amino derivative of MS-27-275 (Saito et al., supra).

F. Other inhibitors, such as: depudecin [its analogues(mono-MTM-depudecin and depudecin-bisether) do not inhibit HDAC] (Kwonet al. 1998. PNAS 95:3356-3361); and scriptaid (Su et al. 2000 CancerResearch, 60:3137-3142).

Compositions

This invention features compositions having the above-describedcompounds. The compositions can further include a second activecompound.

Examples of such a second compound include, but are not limited to, acytokine, a cytokine antagonist, an interleukin, an interleukinantagonist, a growth factor, an angiogenic agent, an anti-histamine, ananti-fibrogenic agent, a vasoactive agent, an antibody, a conjugatedantibody, an adenosine receptor agonist, a peroxisome proliferatingactivator receptor (PPAR) agonist, an anticholinergics, a non-steroidanti-inflammation drug (NSAID), a steroid, an anti-oxidant agent, avitamin, a leukotriene modifier, a mast cell inhibitor, an anti-IgEantibody, lidocaine, epinephrine, a selective serotonin reuptakeinhibitor (SSRI), a 5-hydroxytryptamine (5-HT) receptor antagonist, anantibiotics, a calcineurin inhibitor, an amino acid, a matrixmetalloproteinase (MMP) inhibitor, a DNA methylation inhibitorcollagenase, and clostridium histolyticum.

The compounds described above can be formulated as pharmaceutical orcosmetic compositions.

Pharmaceutical Composition

A pharmaceutical composition of this invention can contain apharmaceutically acceptable carrier, such as a solvent, a dispersionmedium, a coating, an antibacterial and antifungal agent, and anisotonic and absorption delaying agent. A “pharmaceutically acceptablecarrier,” after administered to or upon a subject, does not causeundesirable physiological effects. The carrier in the pharmaceuticalcomposition must be “acceptable” also in the sense that it is compatiblewith the active ingredient and, preferably, capable of stabilizing it.One or more solubilizing agents can be utilized as pharmaceuticalcarriers for delivery of an active compound. Examples of other carriersinclude colloidal silicon oxide, magnesium stearate, cellulose, sodiumlauryl sulfate, and D&C Yellow # 10. The composition can additionallyinclude binding agents (e.g., pregelatinized maize starch,polyvinylpyrrolidone, or hydroxypropyl methylcellulose); binders orfillers (e.g., lactose, pentosan, microcrystalline cellulose, or calciumhydrogen phosphate); lubricants (e.g., magnesium stearate, talc, orsilica); disintegrants (e.g., potato starch or sodium starch glycolate);or wetting agents (e.g., sodium lauryl sulphate). The tablets orcapsules can be coated by methods well known in the art.

Such compositions can be administered orally, parenterally, byinhalation spray, rectally, vaginally, intradermally, transdermally, ortopically in dosage unit formulations containing conventional nontoxicpharmaceutically acceptable carriers, adjuvants, and vehicles asdesired. Topical administration may also involve the use of transdermaladministration such as transdermal patches or iontophoresis devices. Theterm parenteral as used herein includes subcutaneous, intravenous,intramuscular, or intrasternal injection, or infusion techniques.Formulation of drugs is discussed in, for example, Hoover, John E.,Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa.(1975), and Liberman, H. A. and Lachman, L., Eds., Pharmaceutical DosageForms, Marcel Decker, New York, N.Y. (1980).

Cosmetic Composition

Also within the scope of this invention is a cosmetic composition thatcontains one or more active compounds described above. This compositioncontains a safe and effective amount of a dermatologically acceptablecarrier that is suitable for topical application to the skin.

A “cosmetically acceptable” or “dermatologically-acceptable” compositionor component refers a composition or component that is suitable for usein contact with human skin without undue toxicity, incompatibility,instability, allergic response, and the like. It enables an activecompound and optional component to be delivered to the skin at anappropriate concentration(s). The carrier can thus act as a diluent,dispersant, solvent, or the like to ensure that the active materials areapplied to and distributed evenly over the selected target at anappropriate concentration. The carrier can be solid, semi-solid, orliquid. Preferably, it is in the form of a lotion, a cream, or a gel, inparticular one that has a sufficient thickness or yield point to preventthe active materials from sedimenting. The carrier can be inert orpossess dermatological benefits of its own. It should also be physicallyand chemically compatible with the active components described herein,and should not unduly impair stability, efficacy, or other use benefitsassociated with the composition.

A safe and effective amount refers to an amount of a compound,component, or composition sufficient to significantly induce a positivebenefit, preferably a positive skin appearance or feel benefit,including independently the benefits disclosed herein, but low enough toavoid serious side effects, i.e., to provide a reasonable benefit torisk ratio, within the scope of sound medical judgment.

The type of carrier utilized in the cosmetic composition depends on thetype of product form of the composition. A cosmetic composition may bemade into a wide variety of product forms such as those known in theart. These include, but are not limited to, lotions, creams, gels,sticks, sprays, ointments, pastes, and mousses. These product forms maycomprise several types of carriers including, but not limited to,solutions, aerosols, emulsions, gels, solids, and liposomes.

The compositions or preparations for treating a skin disorder orimprovement of skin conditions are generally aimed at providing acondition for increasing skin manageability. There are recognizedcategories of formulations for skin care compositions, including creams,ointments, gels, sprays, lotions, skin tonics, shampoos or mousses asreferred to above Skin sprays are generally composed of aerosolizedcopolymers, such as polyvinylpyrrolidone, vinyl acetate and the like,and may also function as a setting lotion Skin gel preparations aresimilar to sprays in composition, but are in gel and alcohol free form,and can coat the skin. Skin mousse is foam released under pressure froman aerosolized can. The short-chain fatty acid derivative activeingredient in a topical skin care composition according to the presentinvention is preferably present at a concentration of 0.00001 to 100.00%by weight relative to the total weight of the composition, or in adosage of 1 to 1000 mg. A skin care composition for rejuvenating skinaccording to the present invention may be formulated as a hydrophobic orhydrophilic cream, ointment, gel, emollient, spray, lotion, skin tonic,shampoo, body wash or mousse as referred to above, suitably withadditional ingredients suitable for use in skin care compositions oftypes known in the art, and such further ingredients can includepetrolatum, waxes, lanolin, silicone, liposomes, vegetable, mineraloils, plasticizers, fragrances, preservatives, a penetration enhancingagent, a pH adjusting agent or other suitable ingredients for topicalskin compositions. Such ingredients can moisturize skin, stabilize theactive compound, increase drug-skin contact and local concentration,control drug slow release, and/or aid in decreasing skin erythema andbreakage, preventing skin atrophy, fibrosis and infection, and promotingskin wound healing.

Suitable salts for the components to be employed according to thepresent subject matter are also those with inorganic cations, forexample alkali metal salts, in particular sodium, potassium, or ammoniumsalts, alkaline earth metal salts such as, in particular, the magnesiumor calcium salts, as well as salts with bi- or tetravalent cations, forexample the zinc, aluminum, or zirconium salts. Also contemplated aresalts with organic bases, such as dicyclohexylamine salts;methyl-D-glucamine; and salts with amino acids, such as arginine,lysine, histidine, glutamine and so forth. Also, the basicnitrogen-containing groups can be quaternized with such agents as: loweralkyl halides, such as methyl, ethyl, propyl, and butyl chlorides,bromides, and iodides; dialkyl sulfates, such as dimethyl, diethyl,dibutyl, and diamyl sulfates; long chain halides, such as decyl, lauryl,myristyl, and stearyl chlorides, bromides, and iodides; asthma halides,such as benzyl and phenethyl bromides; and others. Salt-forming agents,for example, low molecular weight alkylamines such as methylamine,ethylamine, or triethylamine can also be employed. Water or oil-solubleor dispersible products are thereby obtained.

The amount of active ingredient that can be combined with the carriermaterials to produce a single dosage form will vary depending upon thesubject and the particular mode of administration. The dosage requiredwill vary according to a number of factors known to those skilled in theart, including, but not limited to, the compound or compounds used, thespecies of subject, the size of the subject, and the severity of thesigns and symptoms of the skin. The compounds can be administered in asingle dose, in multiple doses throughout a 24-hour period, or bycontinuous infusion. When administered by continuous infusion, thecompounds can be supplied by methods well known in the art, such as, butnot limited to, intravenous gravity drip, intravenous infusion pump,implantable infusion pump, or any topical routes. Treatment of thesubject with an HDAC inhibitor alone or in combination with other agentsof the invention may last for the life of the subject.

The above-described compositions can contain one or more prodrugs. The“pro-drugs” refers to therapeutic agents or compounds that undergobiotransformation prior to exhibiting their pharmacological effects. Thechemical modification of drugs to overcome pharmaceutical problems hasalso been termed “drug latentiation.” Drug latentiation is the chemicalmodification of a biologically active compound to form a new compoundwhich upon in vivo enzymatic attack will liberate the parent compound.The chemical alterations of the parent compound are such that the changein physiochemical properties will affect the drug's absorption,distribution and enzymatic metabolism. The definition of druglatentiation has also been extended to include nonenzymatic regenerationof the parent compound. Regeneration takes place as a consequence ofhydrolytic, dissociative, and other reactions not necessarily enzymemediated. The terms pro-drugs, latentiated drugs, and bioreversiblederivatives are used interchangeably. By inference, latentiation impliesa time lag element or time component involved in regenerating thebioactive parent molecule in vivo. The term pro-drug is general in thatit includes latentiated drug derivatives as well as those substanceswhich are converted after administration to the actual substance whichcombines with receptors. The term pro-drug is also a generic term foragents which undergo biotransformation prior to exhibiting theirpharmacological actions. In the case where the administered drug is notthe active agent, but rather is biotransformed to the active agent, theterm “HDAC inhibitor” as “pro-drug” also includes compounds which maynot necessarily undergo biotransformation to the administered drug butmay undergo biotransformation to the active agent which exhibits thedesired pharmacological effect.

Methods/Uses

The above-described compositions are particularly useful for improvingaesthetic appearance of skin and for producing smooth skin. Accordingly,this invention features a method of skincare.

The present invention aims at providing methods and compositions havingone HDAC inhibitor or at least one HDAC inhibitor in combination withother compounds to treat a skin disorder or improve aesthetic appearanceof skin, protect skin against stresses, ameliorate wrinkle, producesmooth skin and rejuvenate skin.

The invention is broadly intended for the use of HDAC inhibitors and itspharmaceutically acceptable salts to treat a skin disorder or rejuvenateskin, including, but not limited to, reducing the appearance of linesand/or wrinkles; reducing dermatological signs of chronological aging,photo-aging, hormonal aging, and/or actinic aging; reducing thenoticeability of facial lines, wrinkles and age spots, facial wrinkleson the cheeks, forehead, perpendicular wrinkles between the eyes,horizontal wrinkles above the eyes, and around the mouth, marionettelines, and particularly deep wrinkles or creases; reducing and/ordiminishing the appearance and/or depth of lines and/or wrinkles;improving the appearance of suborbital lines and/or periorbital lines;reducing the appearance of crow's feet; rejuvenating and/or revitalizingskin, particularly aging skin; reducing skin fragility; reducing and/ortreating hyperpigmentation or hypopigmentation, minimizing skindiscoloration; improving skin tone, radiance, clarity and/or tautness;reducing and/or ameliorating skin sagging; improving skin firmness,plumpness, suppleness and/or softness; improving skin texture and/orpromoting retexturization; improving skin barrier repair and/orfunction; improving the appearance of skin contours; restoring skinluster and/or brightness; minimizing dermatological signs of fatigueand/or stress; resisting environmental stress such as weather, sunburn,radiation damage, oxygen radical, hydrogen peroxide, and reactive oxygenintermediates; reducing skin cracking; replenishing ingredients in theskin decreased by aging and/or menopause; increasing cell proliferationand/or multiplication; increasing skin cell metabolism decreased byaging and/or menopause; improving skin moisturization; increasing skinelasticity and/or resiliency; enhancing exfoliation; improvingmicrocirculation; reducing rosacea-associated skin redness; reducingskin erythema, swelling, and crusting; reducing drug-induced skinatrophy; limiting cutaneous deformation; reducing progressivedegradation of a dermal-epidermal junction and/or degradation of acell-cell cohesion in skin; and any combinations thereof, wherein theprogressive degradation of a dermal-epidermal junction and/ordegradation of a cell-cell cohesion in skin is not hyperkeratosis.

In one embodiment, the above-described composition or method is appliedto a subject that is healthy (i.e., a health subject), such as one whois free of cancer or other malignancy. The surface of skin where thecomposition is applied to is free of a wound.

The topical composition is useful for regulating or improving skincondition, including regulating visible or tactile wrinkles ordiscontinuities in skin, e.g., visible and/or tactile wrinkles ordiscontinuities in skin texture or color, more especially thoseassociated with skin ageing. Such wrinkles or discontinuities may beinduced or caused by internal factors (e.g., chronological aging andother biochemical changes from within the skin) or external factors(e.g., ultraviolet radiation, environmental pollution, wind, heat, lowhumidity, harsh surfactants, and abrasives).

Regulating skin conditions can be carried out prophylactically ortherapeutically. Prophylactical regulation includes delaying,minimizing, or preventing the above-mentioned disorder, or visible ortactile wrinkles or discontinuities in skin. Therapeutic regulation, onthe other hand, includes treating the above-mentioned disorder orameliorating, diminishing, minimizing or effacing such wrinkles ordiscontinuities. Regulating skin conditions involves improving skinappearance feel, e.g., providing a smoother, more even appearance, orfeel and reducing signs of aging.

To use a topical composition of this invention, one can topically applyto the skin a safe and effective amount of the composition. The appliedamount, the frequency of application and the period of use vary widelydepending upon the active levels of a given composition and the level oftreatment or regulation desired, e.g., in light of the level of skindisorder or ageing present in the subject and the rate of further skinageing.

A wide range of quantities of the compositions of the present inventioncan be employed to provide a skin appearance and/or feel benefit.Quantities of the compositions typically applied per application arefrom about 0.1 mg/cm² to about 10 mg/cm², e.g., 2 mg/cm². Typically, acomposition can be used once per day. However application rates can varyfrom about once per week up to about three times per day or more.

The topical compositions of this invention provides a visibleimprovement in skin condition essentially immediately followingapplication of the composition to the skin. Such immediate improvementinvolves coverage or masking of skin imperfections such as texturaldiscontinuities (including those associated with skin aging, e.g.,enlarged pores), or providing a more even skin tone or color. Thecompositions of the invention also provide visible improvements in skincondition following chronic topical application. “Chronic topicalapplication” involves continued topical application of a compositionover an extended period during the subject's lifetime, preferably for aperiod of at least about one week, one month, three months, six months,or one year. Chronic regulation of skin condition involves improvementof skin condition following multiple topical applications.

Regulating skin conditions is preferably performed by applying acomposition in the form of a skin lotion, cream, cosmetic, or the likewhich is intended to be left on the skin for an extended period for someaesthetic, prophylactic, therapeutic or other benefit (i.e., a“leave-on” composition). As used herein, “leave-on” compositions excluderinse-off skin cleansing products. After applying the composition to theskin, the leave-on composition is preferably left on the skin for aperiod of at least about 15 minutes, 30 minutes, 1 hour, or up to about12 hours.

This invention also features a method of treating a number of skindisorders. Examples of these disorders include rosacea, acne, pityriasisrosea, inflammatory skin reactions such as urticaria (swelling withraised edges), general swelling, erythema, rhinophyma, acne rosacea,pityriasis rosea, rosacea-associated pimples, papules, pustules, andtelangiectasia. The method includes administering to a subject in needthereof a composition containing an effective amount of theabove-described histone hyperacetylating agent or a pharmaceuticallyacceptable salt thereof, and a cosmetically or pharmaceuticallyacceptable carrier. In one embodiment, the method is used to treat oneor more of the disorders, where no or little itchiness is involved.

Rosacea is a chronic condition characterized by facial erythema orredness. Pimples are sometimes included as part of the definition. Itprimarily affects Caucasians of mainly northwestern European descent,but can also affect people of other ethnicities. Rosacea affects bothsexes, but is almost three times more common in women. It has a peak ageof onset between 30 and 60. Rosacea typically begins as redness on thecentral face across the cheeks, nose, or forehead, but can also lesscommonly affect the neck, chest, ears, and scalp. In some cases,additional symptoms, such as semi-permanent redness, telangiectasia(dilation of superficial blood vessels on the face), red domed papules(small bumps) and pustules, red gritty eyes, burning and stingingsensations, and in some advanced cases, a red lobulated nose(rhinophyma), may develop.

Acne is a common human skin disease, characterized by areas of skin withmultiple noninflammatory follicular papules or comedones and byinflammatory papules, pustules, and nodules in its more severe forms.Acne mostly affects the areas of skin with the densest population ofsebaceous follicles; these areas include the face, the upper part of thechest, and the back. Severe acne is inflammatory, but acne can alsomanifest in noninflammatory forms. Acne lesions are commonly referred toas pimples, blemishes, spots, zits, or simply acne. Acne lesions arecaused by changes in pilosebaceous units, skin structures consisting ofa hair follicle and its associated sebaceous gland, changes whichrequire androgen stimulation. Acne occurs most commonly duringadolescence, affecting more than 89% of teenagers, and frequentlycontinues into adulthood. In adolescence, acne is usually caused by anincrease in male sex hormones, which people of both genders accrueduring puberty. For most people, acne diminishes over time and tends todisappear—or at the very least decrease—after one reaches one's earlytwenties.

Pityriasis rosea is an acute, benign, self-limiting skin rash. Itgenerally begins with a single “herald patch” lesion, followed in 1 or 2weeks by a generalized body rash lasting about 6 weeks. It occurs mostcommonly in people between the ages of 10 and 35, but may occur at anyage. The rash can last from several weeks to several months. Usuallythere are no permanent marks as a result of this condition, althoughsome darker-skinned persons may develop long-lasting flat brown spotsthat eventually fade. It may occur at anytime of year, but pityriasisrosea is most common in the spring and fall. The cause of pityriasisrosea is unknown. It is not a sign of any internal disease, nor is itcaused by a fungus, a bacteria, or an allergy. Recent evidence suggeststhat it may be caused by a virus since the rash resembles certain viralillnesses, and occasionally a person feels slightly ill for a shortwhile just before the rash appears. Pityriasis rosea does not seem tospread from person to person and it usually occurs only once in alifetime.

Urticaria (or hives) is a kind of skin rash notable for dark red,raised, itchy bumps. Hives are frequently caused by allergic reactions;however, there are many non-allergic causes. For example, most cases ofhives lasting less than six weeks (acute urticaria) are the result of anallergic trigger. Chronic urticaria (hives lasting longer than sixweeks) are rarely due to an allergy. The majority of patients withchronic hives have an unknown (idiopathic) cause. Perhaps as many as30-40% of patients with chronic idiopathic urticaria will, in fact, havean autoimmune cause. Acute viral infection is another common cause ofacute urticaria (viral exanthem). Less common causes of hives includefriction, pressure, temperature extremes, exercise, and sunlight.Erythema is a skin condition characterized by redness or rash. There aremany types of erythema, including photosensitivity, erythema multiforme,and erythema nodusum. Photosensitivity is caused by a reaction tosunlight and tends to occur when something, such as an infection or amedication, increases your sensitivity to ultraviolet radiation.Erythema multiforme is characterized by raised spots or other lesions onthe skin. It is usually caused by a reaction to medications, infections,or illness. Erythema nodosum is a form of erythema that is accompaniedby tender lumps, usually on the legs below the knees, and may be causedby certain medications or diseases

A “subject” refers to a human and a non-human animal. Examples of anon-human animal include all vertebrates, e.g., mammals, such asnon-human primates (particularly higher primates), dog, rodent (e.g.,mouse or rat), guinea pig, cat, and non-mammals, such as birds. In apreferred embodiment, the subject is a human. In another embodiment, thesubject is an experimental animal or animal suitable as a disease model.A subject to be treated for the above-described disorder can beidentified by standard diagnosing techniques for the disorder.

“Treating” refers to administration of a compound to a subject, whichhas one of the above-mentioned disorders, with the purpose to cure,alleviate, relieve, remedy, delay the onset of, or ameliorate thedisorder, the symptom of the disorder, the disease state secondary tothe disorder, or the predisposition toward the disorder. An “effectiveamount” refers to an amount of the compound that is capable of producinga medically desirable result, e.g., as described above, in a treatedsubject. The treatment method can be performed alone or in conjunctionwith other drugs or therapy.

The specific examples below are to be construed as merely illustrative,and not limitative of the remainder of the disclosure in any waywhatsoever. Without further elaboration, it is believed that one skilledin the art can, based on the description herein, utilize the presentinvention to its fullest extent. All publications cited herein arehereby incorporated by reference in their entirety. Further, anymechanism proposed below does not in any way restrict the scope of theclaimed invention.

Example 1 Topical Compositions of HDAC Inhibitors A. Preparation of anOleaginous Ointment of Phenylbutyrate:

Sixty-five gram (65 g) of white petrolatum (Riedel-de Haen), 15 g ofcetyl alcohol (Riedel-de Haen), 260 g of soft paraffin (Merck), 155 g ofliquid paraffin (Merck), and 5 g of 4-phenylbutyrate (Merck) were mixedin a beaker and heated at 70° C. to form a paste. The paste was stirredat 400 rpm for 1 hour, and then cooled at room temperature.

B. Preparation of a Cream of Phenylbutyrate:

Part I: 70 g of Tefose 63™, 20 g of Superpolystate™, 10 g of Coster5000™, 15 g of Myriyol 318™, 15 g of Coster 5088™, and 15 g of GMS SE™(all commercially available from a local supplier) were mixed in abeaker and heated at 70° C.

Part II: 5.739 g of sodium 4-phenylbutyrate (Triple Crown America,Inc.), 0.125 g of methylparaben (Merck), 0.075 g of propylparaben(Merck), and 149.061 g of deionized water were mixed in a beaker andheated at 70° C. Part II was slowly added into part I and continuallystirred at 400 rpm for 5 minutes to form a mixture. 2% Stabileze QM™(prepared by dissolving 2 g of Stabileze QM™ in 98 g of deionized water,heating and stirring at 70° C. to form a paste, and cooling at roomtemperature) was added into the mixture and stirred for 5 minutes. ThepH of the mixture was adjusted to 5.34 with 0.85% phosphoric acid(Merck), and stirred at 600 rpm for 20 minutes. The mixture was cooledat room temperature.

C. Preparation of a Gel of Phenylbutyrate:

Part I: 10 g of Stabileze QM™ and 232.035 g of deionized water weremixed in a beaker and heated at 70° C.

Part II: 5.739 g of sodium 4-phenylbutyrate (Triple Crown America,Inc.), 0.125 g of methylparaben (Merck), 0.075 g of propylparaben(Merck), 232.035 g of deionized water, and 20 g of 10% NaOH were mixedin a beaker and heated at 70° C.

Part II was slowly added into part I and continually stirred at 400 rpmfor 20 minutes to form a mixture. The mixture was cooled at roomtemperature.

D: Preparation of a Liposomal Formulation of Phenylbutyrate:

In this liposomal formulation, egg phosphatidylcholine (EPC) andcholesterol were used in equi- or different-molar concentrations asprimary lipid components. Various liposomes located with4-phenylbutyrate were obtained by varying the lipid:phenylbutyrateratio. Liposomes were prepared by thin film hydration, sized by membraneextrusion, and physically evaluated.

E: Preparation of Ointment of Trichostatin A:

To prepare the ointment, 472.5 g of white petrolatum (Riedel-de Haen),27 g of paraffin wax 50/52 (local supplier), and 0.5 g of trichostatin A(sigma) were mixed in a beaker and heated at 70° C. to form a paste. Thepaste was stirred at 400 rpm for 1 hour, and then cooled at roomtemperature.

F. Preparation of an Oleaginous Ointment of Trichostatin A:

To prepare the ointment, 67.5 g of white petrolatum (Riedel-de Haen), 16g of cetyl alcohol (Riedel-de Haen), 260.5 g of soft paraffin (Merck),155.5 g of liquid paraffin (Merck), and 0.5 g of trichostatin A (sigma)were mixed in a beaker and heated at 70° to form a paste. The paste wasstirred at 400 rpm for 1 hour, and then cooled at room temperature.

G. Preparation of Cream of Valproic Acid:

Part I: 70 g of Tefose 63®, 20 g of Superpolystate®, 10 g of Coster5000®, 15 g of Myriyol 318®, 15 g of Coster 5088®, and 15 g of GMS SE.®(all commercially available from local supplier) were mixed in a beakerand heated at 70° C.

Part II: 5.739 g of valproic acid (sigma), 0.125 g of methylparaben(Merck), 0.075 g of propylparaben (Merck), and 149.061 g of deionizedwater were mixed in a beaker and heated at 70° C.

The part II was slowly added into the part I and continually stirred at400 rpm for 5 minutes to form a mixture. 2% Stabileze QM®.D (prepared bydissolving 2 g of Stabileze QM.® in 98 g of deionized water, heating andstirring at 70° C. to form a paste, and cooling at room temperature) wasadded into the mixture and stirred for 5 minutes. The pH of the mixturewas adjusted to 5.34 with 0.85% phosphoric acid (Merck), and stirred at600 rpm for 20 minutes. The mixture was cooled at room temperature.

Example 2

In this example, assays were conducted to show that an HDAC inhibitorwas effective for skin regeneration after abrasive skin damage.

Groups of 5 ICR derived male mice weighing 22±2 g, provided by animalbreeding center of MDS Pharma Service-Taiwan, Ltd., were used. Underhexobarbital (90 mg/kg, IP) anesthesia, the shoulder and back region ofeach animal was shaved. A sharp punch (ID 12 mm) was used to remove theskin including panniculus carnosus and adherent tissues. The wound area,traced onto the clear plastic sheets on day 3, 5, 7, 9, and 11, werequantified by use of an Image Analyzer (Life Science Resources VISTA,Ver. 3.0). The formulation of 5% sodium 4-phenylbutyrate gel or placeboat a dose of 200 μg/mouse was applied topically immediately followinginjury and once daily thereafter for a total of 10 consecutive days. Thewounds half-closure time (CT50) was determined by linear regressionusing Graph-Pad Prism (Graph Pad Software USA) and unpaired Student's ttest was applied for comparison between the phenylbutyrate treated andplacebo group at each measurement time point. Differences wereconsidered statistically significant at p<0.05 (*). As shown in FIG. 1,the phenylbutyrate gel significantly promoted skin regeneration sinceDay 3 (p<0.05).

Example 3

In this example, assays were conducted to show that an HDAC inhibitorproduced smoother skin after skin recovery from radiation damage.

Adult female Sprague Dawley (SD) rats were purchased from the animalcenter of the National Science Council of Taiwan, and weighed 250-300 gat the time of irradiation. Each rat was caged alone and allowed chowand water. They were anesthetized with pentobarbital 50 mg/kg i.p.before irradiation. The skin over the gluteal area was shaved completelyand radiation fields with 2-cm diameter were outlined with a marking penjust prior to irradiation. An electron beam with 6 MeV energy producedby a linear accelerator was used. The dose was delivered on Day 0 at 4Gy/min up to 40 Gy to the prepared area. One group with skin irradiationwas treated by vehicle, another group with skin irradiation was treatedby a 5% phenylbutyrate gel, and the third group with skin irradiationwas left untreated. The vehicle and phenylbutyrate gel were appliedtopically to the irradiated skin twice daily from Day 1 to Day 90 afterirradiation. The mean dosage for each treatment in the respective groupswas 50 mg phenylbutyrate per cm² skin, and an equivalent amount of thevehicle base for the control groups. The irradiated skins were subjectedto H&E histology.

As shown in FIG. 2, the group treated with phenylbutyrate for 180 dayshad smooth skin, thicker epidermis with more cell layers but havethinner dermis (measured from epidermis to the subcutaneous fat layer)with less fragmented collagen deposition when compared to the controlgroups (normal skin and acute reaction on Day 7). In contrast, thevehicle group showed obvious skin wrinkles with more fragmented collagendeposit.

Example 4

Immunofluorescence was conducted to show that an HDAC inhibitorsuppressed TGF-beta, a Fibrogenic Growth Factor, in the late skinremodeling to prevent skin deformation.

The same pathological sections described in Example 3 were subjected toimmunofluorescence with the anti-TGF-beta 1 and 2 antibodies. As shownin FIGS. 3A-3D, the TGF-beta protein, a strong fibrogenic factor, wasup-regulated by irradiation, and highly expressed in fibrogenic skinboth in keratinocytes of the epidermis and in myofibroblasts of thedermis on Day 7 and Day 180 in the acute reaction and vehicle-treatedgroup, respectively. However, the expression of TGF-beta was suppressedeffectively in the phenylbutyrate-treated group on Day 180 which showedless fragmented collagen deposit compared to the control groups.

Example 5

Immunohistochemistry was conducted to show that TNF-alpha, aproinflammatory cytokine, was suppressed by an HDAC inhibitor in thelate skin remodeling to prevent chronic skin ulceration.

On Day 270, three of five rats in Vaseline-treated group and four offive rats in the vehicle-treated group, compared with zero of five ratsin the phenylbutyrate-treated group, showed chronic ulceration,necrosis, bullae formation, and inflammatory cell infiltration. Thedecrease in late radiation-induced skin damage by topical phenylbutyratewas consistent with the suppression of TNF-alpha expression (FIGS.4A-4D).

Example 6

In this example, assays were conducted to show that an HDAC inhibitorpromoted the skin rejuvenation from infection, inflammation, and immunereaction.

Groups of 5 Long Evans rats weighing 150±20 g were used. A well-groundsuspension of killed Mycobacterium tuberculosis (DIFCO, USA; 0.3 mg in0.1 ml of light mineral oil; Complete Freund's Adjuvant, CFA) wasadministered into the subplantar region of the right hind paw. The skinwound was induced by the Mycobacterium tuberculosis injection into thesub-plantar region. The 10% of phenylbutyrate ointment at a dose of 200mg/paw was applied topically twice daily for 18 consecutive days afterbacterial injection. FIGS. 5A and 5B are back views of ankle and plantarjoint. As shown in the figures, skin rejuvenation is promoted in theplantar skin wound in the treated group using phenylbutyrate.

Example 7

In this example, assays were conducted to show that an HDAC inhibitorled to improvement of aesthetic appearance of skin. A 60-year-old femalewith rosacea-associated erythema and papules of the nose and cheeksinitially applied the 2.5% sodium 4-phenylbutyrate gel to her face 2-3times a day. By the third day improvement was evident. Then applicationof the topical gel decreased in frequency to once a day or few times aweek. After several months her face was completely clear ofrosacea-associated redness and papules.

The gel was also used on three other persons who had rosacea (one maleand two female) in the same manner. After the treatment, it was foundthat their faces were also free of rosacea-associated redness andpapules.

Example 8

In this example, assays were conducted to show that an HDAC inhibitorameliorated the tyrosine kinase inhibitor (TKI)-augmented skin reactionin a mouse model.

Tyrosine kinase inhibitors (TKIs) cause acneform-like skin toxicities inhumans. To induce the TKI-augmented skin reaction in a mouse model,groups (n=5, each) of BALB/c male mice weighing 22±2 g received topicalapplication on the ear skin with 10 μL of the solutions of a TKI(PD168393) (4 mmol/L) dissolved in DMSO/absolute ethanol ( 1/10) 30minutes before 10 μL of 0.5% 2,4-dinitrofluorobenzene (DNFB) irritationon the ear of testing animals (Pastore S, et al. J Immunol174:5047-5056, 2005).

To test the therapeutic drug effects on the TKI-augmented skin reaction,a 2.5% sodium 4-phenylbutyrate gel or placebo (gel base) was appliedtopically on the right ear 3 times at 3-hour interval before hand on day0 and day 1. On day 1, 60 minutes after the first dosing ofphenylbutyrate or placebo, the TKI (PD168393) was applied topically 30minutes before 0.5% DNFB irritation on the right ear skin. The secondand third dosing of phenylbutyrate and placebo on day 1 were applied 1hour and 3 hours after DNFB irritation. Ear swelling was measured with aDyer model micrometer gauge at 0, 3, 6, 8, 24 and 48 hours after DNFBirritation. As a treatment control for comparison, the strong steroid,dexamethasone (0.3 mg), was administered topically 60 minutes before and15 minutes after DNFB irritation in a control group. The right and leftear thickness of each mouse was measured with a Dyer model micrometergauge. Ear edema was calculated by subtracting the thickness of the leftear (normal control) from the right ear (treated ear).

Topical administration of 4 mM of the TKI (PD168393) alone, DMSO alone,the placebo gel base only, or the 2.5% phenylbutyrate gel alone had noeffect on ear thickness, and did not induce any change in normal skinhistology. By contrast, 4 mM of the TKI (PD168393) applied 30 minutesbefore DNFB irritation led to aggravation of the DNFB-induced skinresponse. However, the skin pre-treated with the 2.5% phenylbutyrate gelresulted in a significant reduction of the TKI-augmented ear swellinginduced by DNFB irritation at 3, 6, 8, 24, and 48 hours after DNFBirritation as compared to the skin pre-treated with the placebo gel base(FIGS. 6A and 6B). Dexamethasone did not suppress the TKI-augmented skinreaction at 3, 6, and 8 hours after DNFB irritation. Therefore, theseresults suggest that the potent steroid, dexamethasone, is not effectiveon suppression of the TKI-augmented skin reaction, but the 2.5%phenylbutyrate gel appears to have therapeutic benefit on thedermatoloical side effects related to the tyrosine kinase inhibitionwhich will induce acneform skin lesions in humans.

Other Embodiments

All of the features disclosed in this specification may be combined inany combination. Each feature disclosed in this specification may bereplaced by an alternative feature serving the same, equivalent, orsimilar purpose. Thus, unless expressly stated otherwise, each featuredisclosed is only an example of a generic series of equivalent orsimilar features.

A number of embodiments of the invention have been described.Nevertheless, it will be understood that various modifications may bemade without departing from the spirit and scope of the invention.Accordingly, other embodiments are within the scope of the followingclaims.

1. A method for reducing skin wrinkles or treating a skin disorder,comprising administering to a subject in need thereof a compositioncomprising an effective amount of a histone hyperacetylating agent or apharmaceutically acceptable salt thereof, and a cosmetically orpharmaceutically acceptable carrier, wherein the skin disorder isselected from the group consisting of acne and urticaria.
 2. The methodof claim 1, wherein the composition is a topical composition and theadministering step is conducted by applying the composition to a surfaceof skin in need thereof of the subject.
 3. The method of claim 1,wherein the histone hyperacetylating agent is a histone deacetylaseinhibitor.
 4. The method of claim 1, wherein the histonehyperacetylating agent is trichostatin A or trichostatin C.
 5. Themethod of claim 1, wherein the histone hyperacetylating agent isselected from the group consisting of oxamflatin, trapoxin A, FR901228,apicidin, HC-Toxin, WF27082, and chlamydocin.
 6. The method of claim 1,wherein the histone hyperacetylating agent is selected from the groupconsisting of salicylihydroxamic acid, suberoylanilide hydroxamic acid,and azelaic bishydroxamic acid.
 7. The method of claim 1, wherein thehistone hyperacetylating agent is selected from the group consisting ofazelaic-1-hydroxamate-9-an-ilide, M-carboxycinnamic acid bishydroxamide,6-(3-chlorophenylureido)carpoic hydroxamic acid, MW2796, and MW2996. 8.The method of claim 1, wherein the histone hyperacetylating agent isselected from the group consisting of sodium butyrate, argininebutyrate, isovalerate, valerate, 4-phenylbutyrate, sodiumphenylbutyrate, propionate, butrymide, isobutyramide, phenylacetate,3-bromopropionate, valproic acid, valproate, and tributyrin.
 9. Themethod of claim 1, wherein the histone hyperacetylating agent isMS-27-275 or a 3′-amino derivative thereof.
 10. The method of claim 1,wherein the histone hyperacetylating agent is depudecin or scriptaid.11. The method of claim 1, wherein the histone hyperacetylating agent ispresent in an amount of from 0.00001% to 100% by weight of thecomposition.
 12. The method of claim 11, wherein the histonehyperacetylating agent is present in an amount of from 0.0001% to 10% byweight of the composition.
 13. The method of claim 1, wherein thecomposition is a cream, an ointment, a gel, a paste, a powder, anaqueous solution, a spray, a suspension, a dispersion, a slave, alotion, a patch, a suppository, a liposome formation, a mouth wash, anenema, an injection solution, an eye drop, an ear drop, a drip infusion,a microcapsule, or a nanocapsule.
 14. The method of claim 1, wherein thecomposition further comprises a penetration enhancing agent, or a pHadjusting agent to provide a formulation pH in the range ofapproximately 2.0 to 13.0.
 15. The method of claim 1, wherein thecomposition is used as an adjuvant treatment following laser surgery orlaser dermabrasion treatment to enhance aged and injured skinrejuvenation.
 16. The method of claim 1, wherein the method furthercomprises administering to the subject a second agent that comprises acytokine, a cytokine antagonist, an interleukin, an interleukinantagonist, a growth factor, an angiogenic agent, an anti-histamine, ananti-fibrogenic agent, a vasoactive agent, an antibody, a conjugatedantibody, an adenosine receptor agonist, a peroxisome proliferatingactivator receptor (PPAR) agonist, an anticholinergics, a non-steroidanti-inflammation drug (NSAID), a steroid, an anti-oxidant agent, avitamin, a leukotriene modifier, a mast cell inhibitor, an anti-IgEantibody, lidocaine, epinephrine, a selective serotonin reuptakeinhibitor (SSRI), a 5-hydroxytryptamine (5-HT) receptor antagonist, anantibiotics, a calcineurin inhibitor, an amino acid, a matrixmetalloproteinase (MMP) inhibitor, a DNA methylation inhibitor,collagenase, clostridium histolyticum, or combinations thereof.
 17. Themethod of claim 16, wherein the composition and the second agent aresystemically or topically administered simultaneously or sequentially.18. The method of claim 1, wherein the method further comprises, beforethe administering the composition, identifying a subject having thedisorder or skin wrinkles.
 19. The method of claim 1, wherein the methodfurther comprises, after the administering the composition, examiningthe subject for the effect of the composition on the disorder or skinwrinkles.